IGEM:Harvard/2009/Notebook/Harvard iGEM 2010/2010/06/28: Difference between revisions
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[[Image:mrt&ag.jpg|150px|Digest Gel]] | [[Image:mrt&ag.jpg|150px|Digest Gel]] | ||
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!Lane | |||
!Contents | |||
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| Lane 1 | |||
| 1KB Plus Ladder | |||
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| Lane 2 | |||
| pENTCUP2 | |||
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| Lane 3 | |||
| NOSt | |||
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|Lane 4 | |||
| NOSt+STOP | |||
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|Lane 5 | |||
| B15 digested | |||
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|Lane 6 | |||
| B15 undigested | |||
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==Team Genetic Fence== | ==Team Genetic Fence== |
Revision as of 14:47, 28 June 2010
iGEM iGarden | <html><img src="/images/9/94/Report.png" border="0" /></html> Main project page <html><img src="/images/c/c3/Resultset_previous.png" border="0" /></html>Previous entry<html> </html>Next entry<html><img src="/images/5/5c/Resultset_next.png" border="0" /></html> | |||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Team AllergyPCR Reactions Mix of Parts 1,2,3 w/ primers A&B (simultaneous PCR) Annealing Temp: 60C Extension Time: 15 sec 3 reactions (GFP; Bet; LTP)
Step 1: Mix of Parts 1&2 w/ Primers A&2 3 reactions (GFP; Bet; LTP)
Annealing temp: Extension temp: "3 reactions"
Gels 1 (ladder); 2(LTP 1+2); 3 (Bet 1+2); 4(GFP 1+2); 5(Ladder); 6(LTP 1,2,3); 7(Be 1,2,3); 8(GFP 1,2,3); 9(LTP 1,2,3(using .5ul)); 10(GFP 1,2,3 (using .5 uL)) Lanes 2-4 should be ~a little less than 300 bp and 6-10 should be ~450bp Team FlavourMiniprep of Miraculin and Brazzein Constructs
DTL (Digestion, Ligation, Transformation) of The Big Three (pENTCUP2, NOSt, NOSt + STOP)
Team Genetic FenceB21 InnoculationInoculated 3 colonies of B21 transformed E.Coli and set them to shake for 8 hours. We will maxiprep the B21 plasmid for use as a vector backbone. PCR of Gal4DBD and Barnase, attempt 2Ran 3 tubes of each for a total of 6. 7x Mastermix:
Each tube contained:
Reduced annealing temp to 50°C from 56°C which was the last attempt, ran otherwise identical LACIN program Gel of Gal4 DBD and Barnase from second PCR attemptRan 1.2% E-gel of the 6 PCR product tubes.
QIAQuick Purification of PCR ProductBecause colonies 1 and 2 of Barstar worked properly on last week's gel, the PCR products in those two tubes were combined and purified using the Qiagen PCR purification protocol.
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